Toxoplasma is an obligate intracellular parasite that can cause serious disease in immunocompromised patients and in the developing fetus. The majority of Toxoplasma isolates from human patients and livestock in Europe and North-America belong to one of three genetically distinct strains, the type I, type II and type III strains. In mice, these three distinct strains differ enormously in virulence and other phenotypes and there is good evidence that in humans this is also the case. Understanding how these distinct Toxoplasma strains differ in causing disease has important clinical implications; some strains may cause disease through over-stimulation of the immune response leading to toxic levels of cytokines while other strains may cause disease mainly through tissue damage caused by the large numbers of parasites present. If it was known how distinct Toxoplasma strains cause disease and with what strain a patient was infected the treatment could be matched to the specifics of the infection; in some cases inhibition of the immune response might be the best choice while in other cases aggressive treatment with anti-parasitic agents might be needed. Our hypothesis is that, besides host and environmental factors, the genotype of the Toxoplasma strain plays a major role in determining disease outcome through differences between strains in modulating host cell signaling pathways. The goal of this grant is therefore the identification and characterization of Toxoplasma gene products involved in strain-specific modulation of host cell signaling pathways. We will first determine differences between virulent and avirulent strains of Toxoplasma in modulating signaling pathways in mouse innate immune cells in vivo. We will then use existing F1 progeny from crosses between these strains to map the Toxoplasma genomic regions involved. Subsequently, we will use molecular genetic approached to identify the Toxoplasma genes involved. Finally, we will characterize the mechanism of action of these genes. The ability to co-opt host signaling pathways for the parasite's own purposes is likely to be found in other Apicomplexa that are also living within a membrane-limited vacuole, for example, Plasmodium species as they grow within hepatocytes. It is therefore expected that a thorough understanding of the mechanisms Toxoplasma uses to modulate host cell signaling will lead to better therapies against this and other Apicomplexan parasites. PUBLIC HEALTH RELEVANCE: Toxoplasma is an obligate intracellular parasite that can cause serious disease in immunocompromised patients and in the developing fetus. It is expected that the results of this grant will lead to a thorough understanding of the mechanisms Toxoplasma uses to modulate host cell signaling. This could lead to better therapies against this and other Apicomplexan parasites.